KR101828167B1 - Composition for anti-obesity comprising the extract of aronia and method for preparing thereof - Google Patents

Abstract

The present invention relates to a method for extracting aronia to enhance anti-obesity functions in aronia, and an extract thereof. More specifically, the extract, which is obtained by undergoing extraction of aronia involving hot water, ethanol, ultrasound, and super high pressure pulverization-mixing, shows outstanding anti-obesity functions. According to the present invention, the extract increases elution of anti-obesity substances from aronia while improving the anti-obesity functions, thereby ensuring safety and low side effects to human body compared to chemical drugs used to prevent obesity. Therefore, it is possible to remarkably increase value of the aronia extract as food and drug substances.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating obesity, which comprises an extract of Aronia, and a method for preparing the same.

 The present invention relates to a composition for preventing and treating obesity containing an extract of Aronia, and more particularly, to a method for producing Aronia extract, an extract thereof and use thereof for the purpose of enhancing fat-suppressing activity.

Modern society is keeping abundant and convenient life by the development of various technologies. As a result, the proportion of physical activity is reduced and the calories consumed are also reduced compared to the calories consumed. As a result, diseases such as overweight and obesity are increasing worldwide. In Korea, too, the obesity population is rapidly increasing day by day as it gradually becomes a westernized eating habit. In Korea, the ratio of adult obesity is reported to be 37.6% for males and 28.0% for females by 2013, and it is expected to increase further in the future. Obesity has emerged as a serious disease, and the World Health Organization (WHO) has designated obesity as a major health care problem since 2000. Obesity also increases the incidence of complications such as cardiovascular disease, hyperlipidemia, liver dysfunction, diabetes, endocrine disorders, pulmonary dysfunction, and various cancers.

The rapid increase in obesity has created huge profits in the world market for obesity treatment drugs, but the side effects are also very limited, and there are many restrictions on the use of therapeutic drugs. Therefore, it is urgent to develop a therapeutic agent that eliminates side effects of obesity treatment and improves its efficacy. The development of therapeutic agents using natural products has been attracting attention as a way to overcome this. Natural products have been used as medicines for a long time and exhibit numerous physiological activities. Currently, the development of therapeutic agents using natural products is actively under way.

On the other hand, Aronia (Aronia melanocarpa) belongs to Rosaceae and is native to North America and Eastern Europe. Aromania with dark red color has been widely grown all over the world in recent years, and also cultivated as a hybrid breed between two varieties in horticulture.

Aronia is especially well known for its anthocyanin content being among the highest in natural plants. Among the anthocyanins, Cyanidin-3-galactoside is the most abundant and also contains various kinds of anthocyanins. Other physiologically active substances have been studied and reported to contain large amounts of flavonoids and phenols.

It has been shown that these arnia have an excellent antioxidative effect, anti-inflammatory effect and immunity enhancement, and a lot of researches on the anticancer, diabetes and cardiovascular disease-related efficacy.

A common method for extracting natural products is to extract a solvent by using various solvents such as water, ethanol, methanol, and hexane, and further, there is a method of extracting by introducing a physical process such as ultrasonic or ultra-high pressure. have.

However, because anthocyanin is highly unstable in heat, it is urgent to establish an extraction method that can minimize the anthocyanin disruption while enhancing the elution of the active substance for inhibiting obesity in Aronia.

Korean Patent No. 10-1163072

Accordingly, the inventors of the present invention confirmed that it is possible to enhance the obesity-suppressing active substance by various extraction methods in order to maximize the elution of the obesity-inhibiting active substance of Aronia and at the same time to enhance the obesity-suppressing activity.

Accordingly, it is an object of the present invention to provide a method for preparing a composition for preventing and improving obesity, which comprises an extract of Aronia.

Another object of the present invention is to provide a pharmaceutical composition for preventing and treating obesity, which comprises the extract of Aronia according to the present invention as an active ingredient.

It is still another object of the present invention to provide a food composition for prevention and improvement of obesity comprising the extract of Aronia as an active ingredient.

In order to accomplish the above object of the present invention,

(a) lyophilization of Aronia sp. and pulverizing dried Aronia to obtain powder;

(b) mixing the Aronia powder of step (a) with water or ethanol, extracting the mixture using an ultra-high pressure mill, and concentrating and lyophilizing the mixture; And

(c) pulverizing the lyophilized arginia obtained in the step (b) and pulverizing the lyophilized arhonia to obtain a composition for preventing and improving obesity.

In one embodiment of the present invention, the Aromania is selected from the group consisting of black chokeberry, red chokeberry, Aronia melanocarpaarone, Aronia melanocarpaero, Aronia melanoptera, and Aronia melanocarpaquine .

In one embodiment of the present invention, the freezing of step (a) is preferably frozen and frozen at -60 캜 to -50 캜, more preferably frozen at -50 캜, but not always limited thereto.

In one embodiment of the present invention, the extract extracted with water or ethanol is characterized in that it is an extract obtained by treating with an ultra-high pressure milling mixer, not with heat.

In one embodiment of the present invention, the Aromania extract is prepared by mixing (a) Aronia, water, or Aronia with 70% ethanol at a weight ratio of 1:10 (w / w) For 15 minutes; And (b) filtration, concentration under reduced pressure and freeze-drying of the crude extract extracted from the ultra-high pressure milling mixer.

In one embodiment of the present invention, the Aromania extract is prepared by mixing (a) Aronia, water or Aronia with 70% ethanol in a weight ratio of 1:10 (w / w) Lt; RTI ID = 0.0 > 60 C < / RTI > for 12 hours; And (b) filtering and concentrating the extracted Extract and the lyophilization.

In addition, the present invention provides a composition for preventing and improving obesity, which comprises the extract of Aronia produced according to the above method as an active ingredient.

In one embodiment of the present invention, the composition may be a functional food composition having an effect of preventing and improving obesity, or a pharmaceutical composition having an effect of preventing and treating obesity.

The composition for prevention and improvement of obesity, which comprises the Aruonia ultra-high pressure pulverized extract and the ultrasonic extract as an active ingredient, produced according to the production method of the present invention is excellent in the inhibitory activity against obesity and is superior to the chemically synthesized drug It has high safety for human body and low side effects, so it can be useful for functional foods and pharmaceutical materials.

FIG. 1 is a graph showing an evaluation of inhibition of differentiation of adipocytes treated with an extract of Aronia produced according to an embodiment of the present invention.
FIG. 2 is a graph showing the degree of suppression of the expression of PPAR-γ, CREB, SREBP1 and FAS in adipocytes treated with the extract of Aronia produced according to an embodiment of the present invention.
FIG. 3 is a graph showing weight gain of a laboratory rat administered with the extract of Aronia according to an embodiment of the present invention.
FIG. 4 is a photograph of a mouse for experimental administration to which an extract of Aronia was administered according to an embodiment of the present invention. FIG.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for prevention and improvement of obesity, which comprises an Aruonia ultra-high pressure pulverized extract and an ultrasonic extract as an active ingredient.

The composition may be prepared by a method comprising the steps of:

(a) lyophilization of Aronia sp. and pulverizing dried Aronia to obtain powder;

(b) mixing the Aronia powder of step (a) with water or ethanol, extracting the mixture using an ultrahigh-pressure mill or an ultrasonic extractor, concentrating and lyophilizing the mixture; And

(c) pulverizing and lyophilizing the lyophilized arginia obtained in the step (b).

Hereinafter, a method for preparing the Ahnonia extract having the obesity-suppressing activity according to the present invention will be described in more detail.

[Step 1: Aronia  Dryness of life]

The Aronia seeds are placed in a freeze dryer and freeze-dried for 3 days. Specifically, it was dried at -50 ° C and 5 mTorr for 3 days. The dried Aronia is sufficiently pulverized in a pulverizer to prepare a powder.

[Step 2: Aronia  Preparation of Extract Samples]

Extraction is carried out using 10 times by weight water or 70% ethanol solvent for the Aronia powder obtained in the above step 1. Extracts were obtained by passing the extract through an ultra high pressure milling mixer at 20,000 psi pressure for 15 minutes. At this time, it is necessary to use powder which is sufficiently pulverized.

Further, the extraction is carried out using 10 times the weight of water or 70% ethanol solvent for the Aronia powder obtained in the above step 1. Ultrasonic treatment was carried out at 120 kHz for 1 hour and then extracted at 60 ° C for 12 hours to obtain an extract. At this time, it is necessary to use powder which is sufficiently pulverized.

After the extraction is completed, the impurities of the extract are removed using a filter paper, and then the sample is prepared by concentrating the filtrate under reduced pressure and using a freeze dryer.

[Step 3: Aronia  Establishment of Optimal Extraction Method for Improving Obesity Suppression Activity of Extract Samples]

Experiments on obesity inhibitory activity were carried out through the Aronia samples prepared by the method of the present invention, and the optimal extraction process was established after confirming the effect of the ultra high pressure pulverized mixed extract on the obesity inhibitory activity.

In addition, the present invention provides an extract of Aronia with enhanced fat suppressing activity produced by the above extraction method.

The extract of Aronia according to the present invention is prepared by using the above extraction method. In order to avoid the excessive complexity of the specification according to the repetitive description, common description of both inventions is omitted.

The extract of Aronia produced by the method of extracting Aronia according to the present invention is a natural product and has safety and low side effects as a human body and inhibits adipocyte differentiation (see FIG. 1), and induces adipocyte differentiation inducing factor PPAR-γ, CREB, SREBP1, and FAS (see FIG. 2), reducing the body weight gain of experimental rats (see FIGS. 3 and 4) and may be useful in the treatment of obesity.

Accordingly, the composition of the present invention provides a functional food composition having an effect of preventing and improving obesity, or a pharmaceutical composition having the effect of preventing and treating obesity.

In one specific embodiment, the composition of the present invention can be used in the form of a pharmaceutical composition which can be administered orally or parenterally for prevention and improvement of obesity. That is, when the composition of the present invention is used as a pharmaceutical composition, the pharmaceutical composition may be administered in various formulations, oral or parenteral, at the time of actual clinical administration. When the pharmaceutical composition is formulated, it is prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, and surfactants that are generally used. Solid preparations for oral administration are prepared by mixing tablets, pills, powders, granules and capsules. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweetening agents, fragrances and preservatives may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol and gelatin can be used.

In another specific embodiment, the composition of the present invention can be used alone for the improvement of obesity, especially inhibition, or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.

The composition of the present invention can be used in the form of a food composition for prevention and improvement of obesity. When the composition of the present invention is used in the form of a food composition, the food composition may be used as it is, or may be added to other foods or used together with other food or food ingredients. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, the composition of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, it may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient. Such natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like . The ratio of the natural carbohydrate is generally about 0.01 to 0.4 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to these examples.

≪ Example 1 > Preparation of an extract of Aronia

<1-1> Aronia  Ultra-high pressure milling and ultrasonic extraction

100 g of dried Aronia (Aronia melena kappa viking) was mixed with 70% ethanol at a weight ratio of 1:10 (w / w), and then the mixture was subjected to a high-pressure milling mixer at 20,000 psi For 15 minutes, and no heat treatment is performed at the time of extraction. The extracted extracts were filtered and concentrated under reduced pressure, and then lyophilized to obtain powders.

<1-2> Aronia Heat number  Extract preparation ( Comparative Example )

(Arooni melano kappa viking) was freeze-dried for 3 days at -50 ° C and 5 m torr. After freeze-drying, the dried Aronia was sufficiently pulverized in a pulverizer to prepare powder. 100 g of dried Aronia was mixed with 1 L of water and extracted. The hot water extraction was performed at 100 ° C. for 24 hours. The extract was filtered and concentrated through a vacuum filtration apparatus and lyophilized to prepare a powder.

&Lt; 1-3 > Preparation of ethanol extract of Aronia (Comparative Example)

(Arooni melano kappa viking) was freeze-dried for 3 days at -50 ° C and 5 m torr. After freeze-drying, the dried Aronia was sufficiently pulverized in a pulverizer to prepare powder. 100 g of dried Aronia was mixed with 1 L of ethanol and extraction was performed. Ethanol extraction was performed at 80 ° C for 24 hours. The extract was filtered and concentrated through a vacuum filtration apparatus and lyophilized to prepare a powder.

< Experimental Example  1> Aronia  Analysis of cell differentiation inhibition of ultrahigh pressure pulverized mixed extract and ultrasonic extract

In order to confirm the obesity inhibitory activity of the extract of Aronia, the following obesity inhibition experiment was carried out with the extract of Aronia produced in Example 1.

Specifically, the inhibition of adipocyte differentiation of each of the general astrocyte extracts prepared in Examples 1-2 and 1-3 and the ultrasonic pressure-pulverized mixed extracts and ultrasonic extracts prepared in Example 1-1 The culture and differentiation medium were cultured in Dulbecco's modified Eagle medium high glucose supplemented with 10% calf serum and antibiotics at 5% CO ₂ , 37 ° C After confluent cells were incubated with trypsin / EDTA, cells were centrifuged, and the cells were harvested at 6 × 10 5 cells / well at a density of 3 × 10 5 cells / well. (MDI) supplemented with 10% BCS and 23 mg / mL IBMX, 5 mg / mL insulin and 1 mM dexamethasone was induced to induce differentiation for 48 hours, followed by the addition of 5 mg / mL insulin Replace the added medium It gave. The treatment of the sample was carried out from the time of addition of the differentiation inducing medium.

For oil red O staining, 3T3-L1 adipocytes were differentiated for 9 days, then the medium was removed, washed with phosphate buffered saline (PBS), and fixed with 10% formaldehyde solution. After washing with PBS, oil red O solution was added and stained for 30 minutes at room temperature. Oil red O solution was removed, washed with distilled water, and dried. After observation using a phase contrast microscope, And the absorbance at 510 nm was measured.

As a result, the Aroma ultrahigh pressure pulverized mixed extract and the ultrasonic extract increased the activity of inhibiting adipocyte differentiation compared to the general Aroma extract. The Aroma ultrahigh pressure pulverized mixed extract and the ultrasonic extract Increased the activity of inhibiting the differentiation of adipocytes 3T3-L1 (see FIG. 1).

< Experimental Example  2> Aronia  Adipocyte differentiation of ultrasound-pulverized mixed extract and ultrasonic extract Induction factor  Expression analysis

In order to analyze the induction factors of adipocyte differentiation induction factors of Aroma ultrahigh pressure pulverized mixed extract and ultrasound extract, Arnea ultrahigh pressure pulverized mixed extract, Aroniae hot water extract, Aronia hot water extract and Aronia ethanol extract were applied to lipid precursors .

(50, 100, 200, and 400 ug / ml) of Arnoia ultrahigh pressure pulverized mixed extract, Aronia ultrasound extract, Aronia hot water extract and Aronia ethanol extract were added to 6 well plates in which lipogenic precursor cells 3T3- 3T3-L1 cells cultured for 3 days were washed with PBS and homogenized using RIPA buffer. Then, the cells were separated by 10% sodium dodecyl sulfate (SDS) -PAGE and transferred to PVDF membrane. The primary antibodies, PPAR-γ, CREB, SREBP1, and FAS (PPAR-γ, CREB, FAS, cellsignaling, Danvers, MA, USA; SREBP1, Abcam, Cambridge , MA, USA) was reacted at 4 ° C for 12 hours, and the secondary antibody polymerized with HRP was reacted. Immuno-positive bands were developed by enhanced chemiluminescent and chemiluminescent detection systems.

As a result, it was found that the Aroma ultrahigh pressure pulverized mixed extract and the ultrasonic wave extract were superior to the Aronia hot water extract and Aronia ethanol extract PPAR-γ, CREB, SREBP1 and FAS, and the inhibitory effect was confirmed in the ultrasonic extract (see FIG. 2).

< Experimental Example 3> Ultra high pressure  Grinding blend Aronia  Analysis of obesity inhibitory activity of extracts

In order to confirm the obesity inhibitory activity of the extract of Aronia, the body weight change was confirmed by taking the Aronia super -

Specifically, the experimental mice (C57BL / 6N, male, 5 weeks old) were divided into the basic diet (solid diet, Cargill Agri Purina, Kunsan, Korea / 10) were assigned to each group so as to have a similar average weight after each group. The control diet was supplemented with 10 kcal, high-fat diets were consumed at 60 kcal, and high-fat diets were consumed at 100 mg / kg, 200 mg / kg and 400 mg / kg. Water was free-fed and weighed at weekly intervals.

The control diet (10 kcal% fat) High fat diet (60 kcal% fat) g kcal g kcal Casein, 80 Mesh 200 800 200 800 L-Cystine 3 12 3 12 Corn starch 315 1,260 0 0 Maltodextrin 10 35 140 125 500 Sucrose 350 1,400 68.8 275 Cellulose 50 0 50 0 Soybean oil 25 225 25 225 Lard * 20 180 245 2,205 Mineral mix 10 0 10 0 Dicalcium phosphate 13 0 13 0 Calcium carbonate 5.5 0 5.5 0 Potassium citrate, 1H ₂ O 16.5 0 16.5 0 Vitamin mix 10 40 10 40 Choline bitartrate 2 0 2 0 Total 1,055 4,057 773.8 4,057

As a result, the body weight gain of the experimental rats administered with the Aroonia ultra high pressure pulverized mixed extract was confirmed, and the weight increase effect was reduced at all the concentrations compared to the high fat diet. At this time, until the end of 2 weeks, the best effect was observed at a dose of 100 mg / kg, which was the best, and at 3 weeks, 200 and 400 mg / kg, ).

In addition, when the experimental rats administered with the Aroma ultra high-pressure pulverized mixed extract were visually examined, it was found that the high-fat diet was larger than that of the control diet, and that the high fat diet was reduced in the rats fed with the Aaroniae extract The mass was verified by naked eyes, and the effect of inhibiting the obesity of the Aroma ultra high-pressure pulverized mixed extract was confirmed (see FIG. 4).

Therefore, the extract of Aronia produced by the method of extracting Aronia according to the present invention inhibits adipocyte differentiation, inhibits the expression of PPAR-γ, SREBP1, CREB and FAS which are adipocyte differentiation inducers, Thus, it can be effectively used for the treatment of obesity-related diseases.

The present invention has been described with reference to the preferred embodiments and experimental examples. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (9)

A method for preparing a composition for preventing and improving obesity,
The above-
(a) Rapidly frozen at -50 ° C and freeze-dried for 3 days at 5 mTorr to obtain dried powder;
(b) mixing the Aronia powder of step (a) with 70% ethanol at a weight ratio of 1:10 (w / w), passing through an extract for 15 minutes under a pressure of 20,000 psi using an ultra- To obtain an extract;
(c) concentrating and freeze-drying the extract; And
(d) pulverizing and lyophilizing the lyophilized extract obtained in the step (c)
In the extraction step (b), without any heat treatment,
The composition increases the cell differentiation inhibitory activity of adipocyte 3T3-L1,
Wherein the composition inhibits the expression of PPAR-gamma, CREB, SREBP1 and FAS which are inducers of differentiation of adipocytes. The method according to claim 1,
Wherein said Aronia is selected from the group consisting of black chokeberry, red chokeberry, Aronia melanocarpaarone, Aronia melanoptera, Aronia melanoptera, and Aronia melanocarpaekinji. A method for preparing a composition for improvement. delete delete delete delete delete A pharmaceutical composition for the prevention and treatment of obesity comprising a composition produced by the method of claim 1. A food composition for preventing and improving obesity comprising a composition produced by the method of claim 1.

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